Reaction mass of 2-hydroxy-1,3-propanediyl bismethacrylate and 101525-90-0

Endpoint summary

Description of key information

Skin irritation

no data available for GMDA

Read across from analogous substances (see additional information and category document, chapter 5.3)

EGDMA (closely analogous category member): slightly irritant after 24 h occlusive conditions (FDA Draize test, non-GLP; IBR, 1977a)

HPMA (analogous substance of the primary metabolites of GDMA): not irritant after 24 h occlusive conditions (FDA Draize test, non-GLP; IBR, 1977b)

Eye irritation

GDMA: Irritating to the Human Cornea in vitro (OECD 492, GLP; Envigo 2018)

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Reference

Reference 1

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Study well documented, meets generally accepted scientific principles, acceptable for assessment.

Qualifier:

according to guideline

Guideline:

other: Appraisal of the safety of Chemicals in foods, drugs and cosmetics by staff of the Division of Pharmacology, FDA acc. to Draize

Principles of method if other than guideline:

Method: Appraisal of the safety of chemicals in foods, drugs and cosmetics by the Staff of the Division of Pharmacology, FDA, Hautgiftigkeit nach Draize (1959)

GLP compliance:

no

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: no data
- Age at study initiation: no data
- Weight at study initiation: 2.5 kg (average)
- Housing: individual cages
- Diet: rabbit standard diet (Höing 222)
- Water ad libitum


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 -1 (max. limitation)
- Humidity (%): 50-60
- Photoperiod ( hrs light): 12

Type of coverage:

occlusive

Preparation of test site:

other: shaved, shaved and scarified

Vehicle:

unchanged (no vehicle)

Controls:

other: as a contol 2 areas of the treated animals were also shaved ans scrarified but remained untreated

Amount / concentration applied:

TEST MATERIAL
- Amount applied (volume): 0.5 ml

Duration of treatment / exposure:

24 hour(s)

Observation period:

72 h

Number of animals:

6

Details on study design:

TEST SITE
- Area of exposure: 2.5 x 2.5 cm
- Type of wrap if used: rubberized cloth

CONTROL.
- as a contol 2 areas of the treated animals were also shaved and scrarified but remained untreated

REMOVAL OF TEST SUBSTANCE
- Washing : no wahing was done


SCORING SYSTEM:
1. Erythema and scars formation
no erythema 0
very slight erythema 1
clear erythema 2
moderate to severe erythema 3
severe erythema (scarlet red) 4
and slightly scars formation

2. Edema formation
no edema 1
very slight edema 2
moderate edema (thickness ca. 1mm) 3
severe edema (thickness more than 4
1mm, large than the edge of the
contact)

Irritation parameter:

erythema score

Basis:

animal: #1, #2, #3, #4, #5, #6

Time point:

other: 24/72h

Score:

0

Max. score:

4

Irritation parameter:

edema score

Basis:

animal: #1, #2, #3, #4, #5, #6

Time point:

other: 2472h

Score:

0

Max. score:

4

 

Animal Nr.			Scores (24 h)	Scores (72 h)
1	shaved	Erythema and scras formation	0	0
		Edema formation	0	0
	shaved/scarified	Erythema and scras formation	0	0
		Edema formation	0	0
2	shaved	Erythema and scras formation	0	0
		Edema formation	0	0
	shaved/scarified	Erythema and scras formation	1	1
		Edema formation	1	1
3	shaved	Erythema and scras formation	0	0
		Edema formation	0	0
	shaved/scarified	Erythema and scras formation	1	0
		Edema formation	1	0
4	shaved	Erythema and scras formation	0	0
		Edema formation	0	0
	shaved/scarified	Erythema and scras formation	0	0
		Edema formation	0	0
5	shaved	Erythema and scras formation	0	0
		Edema formation	0	0
	shaved/scarified	Erythema and scras formation	0	0
		Edema formation	0	0
6	shaved	Erythema and scras formation	0	0
		Edema formation	0	0
	shaved/scarified	Erythema and scras formation	1	1
		Edema formation	0	0

 

1. Primary irritation index of the shaved skin

 

Animal No.	Primary irritation index
1	0.00
2	0.00
3	0.00
4	0.00
5	0.00
6	0.00
X1	0.00

 

 

2. Primary irritation index of the shaved/scarifized skin

 

Animal No.	Primary irritation index
1	0.00
2	1.00
3	0.50
4	0.00
5	0.00
6	0.50
X2	0.33

 

Total index: X₁ +   X₂= 0.33

 

 

• = fragmentary value

 

 

Reevaluation according to OECD 404

 

	Erythema		Edema
	24h	72h	24h	72h
Tier 1	0	0	0	0
Tier 2	0	0	0	0
Tier 3	0	0	0	0
Tier 4	0	0	0	0
Tier 5	0	0	0	0
Tier 6	0	0	0	0
Mean	0	0	0	0
Total mean	0		0
	Erythema		Edema

 

 

Interpretation of results:

not irritating

Remarks:

Migrated information Criteria used for interpretation of results: other: EU-GHS

Conclusions:

Classification: not irritating

Executive summary:

2 -Hydroxypropyl methacrylate was tested in a primary skin irritation test to rabbits according to "Appraisal of the safety of chemicals in food, drugs and cosmetics by the Staff of the Division of Parmacology, FDA, 1959 in food, drugs and cosmetics". .Therefore Hydroxypropyl methacrylate is not irritating according to EU-CLP requirements.

Reference 2

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

test procedure in accordance with national standard methods with acceptable restrictions

Remarks:

Pre-guideline study but comparable to guideline study. Test procedure in accordance with national standard methods with acceptable restrictions. Restrictions: Observation period only 72 h, only two observations, duration of treatment 24 h instead of 4h.

Qualifier:

according to guideline

Guideline:

other: according to Appraisal of the Safety of Chemicals in foods, drugs and cosmetics, FDA Draize (1959)

Principles of method if other than guideline:

Method: according to Appraisal of the Safety of Chemicals in foods, drugs and cosmetics, FDA Draize (1959)
Information about the method, described in the study report ( Draize test: skin irritation/corrosion), is given in the free text field "methods and
materials".

GLP compliance:

no

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: no data
- Age at study initiation: no data
- Weight at study initiation: mean value 2,5 kg
- Housing: single housing
- Diet: ad libitum
- Water: ad libitum
- Acclimation period: no data


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 ±1°C
- Humidity (%): 50 - 60 %
- Air changes (per hr): no data
- Photoperiod (hrs dark / hrs light): 12 hrs dark / 12 hrs light

Type of coverage:

occlusive

Preparation of test site:

other: shaved and shaved/abraded

Vehicle:

unchanged (no vehicle)

Controls:

other: Untreated skin areas of the test animals serve as the control (shaved, scarified).

Amount / concentration applied:

undiluted 0.5 mL

Duration of treatment / exposure:

24 hour(s)

Observation period:

24h and 72h post application

Number of animals:

6

Irritation parameter:

primary dermal irritation index (PDII)

Basis:

mean

Time point:

other: 24 and 72 hours

Score:

0.42

Max. score:

8

Reversibility:

fully reversible

Remarks on result:

other: Only data of shaved skin have been used, data of shaved and scarified skin have not been used.

Irritation parameter:

erythema score

Basis:

mean

Time point:

other: 24 and 72 hours

Score:

0.42

Max. score:

4

Reversibility:

fully reversible

Remarks on result:

other: Only data of shaved skin have been used, data of shaved and scarified skin have not been used.

Irritation parameter:

edema score

Basis:

mean

Time point:

other: 24 and 72 hours

Score:

0

Max. score:

4

Reversibility:

fully reversible

Remarks on result:

other: Only data of shaved skin have been used, data of shaved and scarified skin have not been used.

Overall primary irritation score (PDII): 0.42 of 8 scores FDA (Draize), 1959,  

re-evaluated according to OECD 404

Reevaluation of the test results according to OECD404/GHS: only records for the shaved skin (not scarified) were considered  Erythema/ 24h     Erythema/72h  Oedema/24h  Oedema/72h  animal 1  1  0  0  0  animal 2  0  0  0  0  animal 3  1  0  0  0  animal 4  1  0  0  0  animal 5  1  0  0  0  animal 6  1  0  0  0  average (single scores: animal 1-6)  0,8333  0  0  0  Primary irritation index I = Mean erythema score  (Erythema; 24h, 72h);  out of 4 scores  0,42  Primary irritation index II =  Mean oedema score (Oedema; 24h, 72h);   out of 4 scores  0    Overall average PDII out of 8 scores; For evaluation according to GHS classification system:  0,42

Remarks concerning the study result: The study for acute skin irritation/ corrosion was performed before OECD 404 came into force. For this reason the 
test values were reevaluated according to OECD criteria and test scores (erythema, oedema) obtained for the scarified skin were regarded as irrelevant.

Interpretation of results:

GHS criteria not met

Conclusions:

Ethylene glycol dimethacrylate was not irritating in a primary skin irritation study with in rabbits (24/72-hour occlusive application, no wash of the test substance, re-evaluated according to OECD 404).

Executive summary:

In a primary pre-guideline dermal irritation study New Zealand White rabbits were dermally exposed (intact and scarified skin) to 0.5 mL undiluted Ethyleneglycol dimethacrylate for 72 hours. Animals then were observed for 3 days. Irritation was scored by the method of Draize et al, 1959.

The mean erythema score (average value of  the single scores (animals 1-6; erythema; intact skin, 24h and 72h)  was determined to be 0.42 out of 4 and the mean edema score 0 out of 4. Therefore Ethylene glycol dimethacrylate is

not a dermal irritant .

Therefore the test substance has not to be classified - according to GHS classification criteria and according to Draize-criteria - as non irritant for skin (GHS-hazard category: none).

Remarks concerning the study result: The study for skin irritation/ corrosion was performed before OECD 404 came into force. For this reason the test values were reevaluated according to OECD criteria and test scores (erythema, oedema) obtained for the scarified skin were regarded as irrelevant.

NOTE: Any of data in this dataset are disseminated by the European Union on a right-to-know basis and this is not a publication in the same sense as a book or an article in a journal. The right of ownership in any part of this information is reserved by the data owner(s). The use of this information for any other, e.g. commercial purpose is strictly reserved to the data owners and those persons or legal entities having paid the respective access fee for the intended purpose.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Eye irritation

Link to relevant study records

Reference

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

17.08.2017 to

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 492 (Reconstructed Human Cornea-like Epithelium (RhCE) Test Method for Identifying Chemicals Not Requiring Classification and Labelling for Eye Irritation or Serious Eye Damage)

Version / remarks:

Version July 2015

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: MatTek Corporation Protocol: EpiOcular™ Eye Irritation Test (OCL-200-EIT) for the prediction of acute ocular irritation of chemicals; for use with MatTek Corporation’s Reconstructed Human EpiOcular™ Model; 29 June 2015.

Version / remarks:

Version 29.06.2015

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

Batch: 1270300006
Purity: Reactiveester content: 95.3%
Diester content (isomer mix): 86.4%
Methacrylic acid: 0.86%
Appearance: Colourless to yellowish, liquid
Expiry Date: 01 February 2018
Storage Conditions: At room temperature

Species:

other: EpiOcular Kit and MTT-100

Details on test animals or tissues and environmental conditions:

EpiOcular™ kits and MTT-100 kits are purchased from MatTek Corporation (82105 Bratislava, Slovakia). The EpiOcular™ tissue consists
of normal, human-derived epidermal keratinocytes which have been cultured to form a stratified squamous epithelium similar
to that found in the human cornea. It consists of highly organized basal cells which progressively flatten out as the apical surface
of the tissue is approached, analogous to the normal in vivo corneal epithelium. The EpiOcular™ tissues (surface 0.6 cm²)
were cultured on specially prepared cell culture inserts (MILLICELL(R), 10 mm ).
EpiOcular™ tissues were shipped at 2 - 8 °C on medium-supplemented agarose gels in a 24-well plate.
On day of receipt (19 September 2017) of the EpiOcular™ tissues, the equilibration step
(15 minutes at room temperature in the 24-well shipping container) started. 1.0 mL of the medium was aliquoted into the appropriate wells of pre-labelled 6-well plates.

Controls:

yes, concurrent positive control

yes, concurrent negative control

Amount / concentration applied:

50 µL

Duration of treatment / exposure:

30 min

Duration of post- treatment incubation (in vitro):

120 min

Number of animals or in vitro replicates:

2

Details on study design:

After the overnight incubation, the tissues were pre-wetted with 20 μL of Ca++Mg++free-DPBS. The tissues were incubated at standard culture conditions (37 ± 1.5 °C, 5 ± 0.5% CO2, 95% RH) for 30 minutes.
Test item exposure: After the 30 minute Ca++Mg++free-DPBS pre-treatment, the test and control items were tested by applying 50 μL topically on the EpiOcular™ tissues. The tissues were incubated at standard culture conditions for 30 minutes.
At the end of the 30 minutes treatment time, the test item was removed by extensively rinsing the tissues with Ca++Mg++-free DPBS (brought to room temperature). Three clean beakers containing a minimum of 100 mL each of Ca++Mg++-free DPBS were used per test item. Each test item utilized a different set of three beakers. The inserts containing the tissues were lifted out of the medium by grasping the upper edge of the plastic "collar" with fine forceps. To assure throughput, the tissues were rinsed two at a time by holding replicate inserts together by their collars using forceps. The test or control items were decanted from the tissue surface onto a clean absorbent material and the cultures dipped into the first beaker of DPBS, swirled in a circular motion in the liquid for approximately 2 seconds, lifted out so that the inserts were mostly filled with DPBS, and the liquid was decanted back into the container. This process was performed two additional times in the first beaker. The culture was then rinsed in the second and third beakers of DPBS three times each in the same fashion. Finally, any remaining liquid was decanted onto the absorbent material by rotating the insert to approximately a 45° angle (open end down) and touching the upper lip to the absorbent material (to break the surface tension).
After rinsing, the tissues were immediately transferred to and immersed in 5 mL of previously warmed Assay Medium (room temperature) in a pre-labelled 12-well plate for a 12 minute immersion incubation (post-soak) at room temperature. This incubation in Assay Medium was intended to remove any test item absorbed into the tissue.
At the end of the post-soak immersion, each insert was removed from the Assay Medium, the medium was decanted off the tissue, and the inserts were blotted on absorbent material, and transferred to the appropriate well of the pre-labelled glass tube containing 1 mL of warm Assay Medium. The tissues were incubated for 120 minutes at 37 ± 1.5 °C in a humidified atmosphere of 5 ± 0.5% CO2 (post-treatment incubation).

MTT Assay
After post-treatment incubation of 120 minutes, the MTT assay was performed.
At the end of the post-treatment incubation, each insert was removed from the 6-well plate and gently blotted on absorbent material. The tissues were placed into the 24-well plate containing 0.3 mL of MTT solution. Once all the tissues were placed into the 24-well plate, the plate was incubated for 180 minutes at standard culture conditions.
Each insert was removed from the 24-well plate after 180 minutes, the bottom of the insert was blotted on absorbent material, and then transferred to a pre-labelled 24-well plate containing 2.0 mL of isopropanol in each designated well so that isopropanol was flowing into the insert on the tissue surface. The plates were sealed with a standard plate sealer, and were stored overnight at 2-8 °C in the dark and then shaken for 2-3 hours at room temperature. At the end of the extraction period, the tissue was pierced and the liquid within each insert was decanted into the well from which it was taken.
The extract solution was mixed and two 200 μL aliquots were transferred to the appropriate
wells of a pre-labelled 96-well plate.
The absorbance at 570 nm (OD570) of each well was measured with a plate reader (Versamax® Molecular Devices, 85737 Ismaning, Germany, Software Softmax Pro Enterprise, version 4.7.1). No reference wavelength measurement was used.

Irritation parameter:

other: Mean tissue viability

Remarks:

Mean relative absorbance [%]

Run / experiment:

Mean of duplicate test

Value:

45.3

Negative controls validity:

valid

Remarks:

100 %

Positive controls validity:

valid

Remarks:

36.3 %

Remarks on result:

positive indication of irritation

Other effects / acceptance of results:

Prediction Model
If the test item-treated tissue viability is > 60% relative to the negative control treated tissue viability, the test item is labeled non-irritant (no Category according to UN GHS).
If the test item-treated tissue viability is ≤ 60% relative to negative control treated tissue viability, the test item is labeled irritant (Category 2 or Category 1 according to UN GHS; no differentiation between the categories possible).

Acceptability of the Assay
The results are acceptable according to MatTek Protocol, if:
1) The negative control OD is > 0.8 and < 2.5,
2) The mean relative viability of the positive control is below 50% of the negative control viability.
3) The difference of viability between the two relating tissues of a single test item is < 20% in the same run (for positive and negative control tissues and tissues of test items). This applies also to the freeze-killed tissues (items and negative control) and the additional viable tissues (without MTT addition) which are calculated as percent values related to the viability of the relating negative control.

Results after treatment for 30 minutes with Glycerol dimethacrylate and the controls

Dose Group	Ab-sorbance Well 1 (Tissue 1/2)	Ab-sorbance Well 2 (Tissue 1/2)	Mean Absor-bance (Tissue 1/2)	Mean Absorbance* Tissue 1 and 2	Mean Absorbance of 2 Tissues*	Rel. Absorbance [%] Tissue 1 and 2**	Absolute Value of the Difference of the Rel.Absorbances [%] Tissue 1 and 2	Mean Rel. Absorbance [%]***
Blank	0.036	0.036	0.036
Negative Control	1.628	1.644	1.636	1.601	1.590	100.6	1.3	100.0
	1.617	1.614	1.616	1.580		99.4
Positive Control	0.585	0.635	0.610	0.574	0.577	36.1	0.3	36.3
	0.618	0.613	0.615	0.580		36.4
Test Item	0.694	0.744	0.719	0.683	0.721	43.0	4.7	45.3
	0.787	0.800	0.793	0.758		47.6

 

* Mean of two replicate wells after blank correction

** Relative absorbance [rounded values]

 

*** Mean relative absorbance [rounded values]

The optical pre-experiment (colour interference pre-experiment) to investigate the test item’s colour change potential in water or isopropanol did not lead to a change in colour.

Optical evaluation of the MTT-reducing capacity of the test item with MTT-reagent did not show blue/purple colour.

The mean relative absorbance value of the test item, corresponding to the cell viability, decreased to 45.3% (threshold for irritancy: ≤ 60%), consequently the test item was irritant to eye.

Concerning acceptance criteria:

• The negative control OD is > 0.8 and < 2.5 (1.614 and 1.644).

• The mean relative viability of the positive control is below 50% of the negative control viability (36.3%).

• The difference of viability between the two relating tissues of a single item is < 20% (values between 0.3% and 4.7%) in the same run (for positive and negative control tissues and tissues of single test items).

Interpretation of results:

Category 2 (irritating to eyes) based on GHS criteria

Conclusions:

It can be stated that in this in vitro GLP study and under the experimental conditions reported, Glycerol dimethacrylate possesses an eye irritating potential.

Executive summary:

An in vitro study was performed to assess the eye irritation potential of Glycerol dimethacrylate by means of the Human Cornea Model Test according to OECD 492 and current GLP requirements.

Additional tests with viable or freeze-killed tissues were not performed, since the test item was not coloured intensively, did not dye water or isopropanol, and did not prove to be a MTT reducer.

Tissues of the human cornea model EpiOcular™ were treated with the test item, the positive and the negative control for 30 minutes each in duplicate.

50 µL of the test item and of the controls, respectively, were applied to each tissue, spread to match the tissue size.

Treatment with the positive control induced a decrease in the mean relative absorbance compared with the negative control to 36.3%, thus the validity of the test system is ensured.

Relevant irritating effects were observed following 30 minutes incubation with Glycerol dimethacrylate. The mean relative absorption value of the tissues corresponding to the cornea viability decreased to 45.3% compared with the value of the negative control (threshold for irritancy: ≤ 60%).

In conclusion, it can be stated that in this study and under the experimental conditions reported, Glycerol dimethacrylate possesses an eye irritating potential.

A limitation of the Test Guideline OECD 492 is that it does not allow discrimination between eye irritation/reversible effects on the eye (Category 2) and serious eye damage/irreversible effects on the eye (Category 1), nor between eye irritants (optional Category 2A) and mild eye irritants (optional Category 2B), as defined by UN GHS.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Skin irritation

No study is available for GMDA.

Read across evaluation according to ECHA’s Read Across Assessment Framework (RAAF)

It is postulated that MAA as common primary metabolite of all category members including GDMA has the strongest impact on local effects driven by its acidic properties. The assessment for the few data gaps for category members can be made by interpolation from other category members. Slight differences in both skin and eye irritation between the category members can be interpreted as patterns according to RAAF nomenclature. However the differences are weak in terms of effect strength and generally below classification tresholds for both endpoints (with exception of GDMA for eye irritation). Considering the observed maximum mild local effects of the category members (again, with exception of GDMA), Scenario 6 of the RAAF approach is considered the most suitable one for the assessment.

There are data for all but one member of this category, GDMA. However, even after 24h contact the studied category members showed only slight irritation potential at most. By interpolation within the series, namely EGDMA, plus in vivo data from HPMA (Hydroxypropyl Methacrylate) as analogous substance to the primary metabolites of GMDA, GDMA itself is considered as slightly irritating.

Eye irritation

An in vitro study was performed to assess the eye irritation potential of GDMA by means of the Human Cornea Model Test according to OECD 492 and current GLP requirements.

Additional tests with viable or freeze-killed tissues were not performed, since the test item was not coloured intensively, did not dye water or isopropanol, and did not prove to be a MTT reducer.

Tissues of the human cornea model EpiOcular™ were treated with the test item, the positive and the negative control for 30 minutes each in duplicate.

50 µL of the test item and of the controls, respectively, were applied to each tissue, spread to match the tissue size.

Treatment with the positive control induced a decrease in the mean relative absorbance compared with the negative control to 36.3%, thus the validity of the test system is ensured.

Relevant irritating effects were observed following 30 minutes incubation with GDMA. The mean relative absorption value of the tissues corresponding to the cornea viability decreased to 45.3% compared with the value of the negative control (threshold for irritancy: ≤ 60%).

In conclusion, it can be stated that in this study and under the experimental conditions reported, GDMA possesses an eye irritating potential.

A limitation of the Test Guideline OECD 492 is that it does not allow discrimination between eye irritation/reversible effects on the eye (Category 2) and serious eye damage/irreversible effects on the eye (Category 1), nor between eye irritants (optional Category 2A) and mild eye irritants (optional Category 2B), as defined by UN GHS. Due to the mild effect strength seen in other studies with category substances, serious eye damage/irreversible effects on the eye can be excluded for GDMA with a high level of confidence so that a Cat 2 classification according to CLP/UN-GHS is considered as sufficient to describe this hazard.

Justification for classification or non-classification

Based on a read across assessment, GDMA is classified as not a skin irritant according to Annex I of CLP/EU-GHS (1272/2008/EC) and UN-GHS.

Based on a valid in vitro study, GDMA

is classified as eye irritant Cat 2 according to Annex I of CLP/EU-GHS (1272/2008/EC) and

as eye irritant Cat 2B according to

UN-GHS.